多药抗性是非小细胞肺癌化疗固有的弱点，全球普遍存在。在临床上，人参和包含人参的中药处方通常用作抗肿瘤辅助治疗，因其具有益气的特性，可以提高化疗药物的疗效并减轻其毒副作用。三萜皂苷是人参中关键的有效成分，人参皂苷Rb1含量最高。然而，人参皂苷Rb1对肿瘤耐药逆转作用和机制的研究仍不明确。本研究旨在系统评价人参皂苷Rb1对A549/DDP细胞铂类药物不敏感的逆转作用并揭示其潜在的分子机制。利用MTT实验评估人参皂苷Rb1在体外对A549/DDP细胞铂类药物不敏感的逆转作用，并通过建立BALB/c-nu小鼠皮下移植肿瘤模型进行研究。此外，还采用P-gp ATP酶活性测定、铂类药物积累实验、Annexin V-FITC凋亡实验、实时荧光定量PCR分析和蛋白质印迹分析等方法来澄清潜在的机制。人参皂苷Rb1可以有效地逆转A549/DDP细胞体内外对铂类药物的抗药性。在人参皂苷Rb1加铂类药物的联合治疗后，A549/DDP细胞中铂类药物的积累增加，同时ABCB1、SHH、PTCH1和GLI2的mRNA和蛋白质表达水平下调，铂类药物诱导的凋亡能力通过对Bax和Bcl-2蛋白质表达水平的相关调节得到提高。更为重要的是，CYP3A4 mRNA和蛋白质水平的变化不显著。人参皂苷Rb1可以增加A549/DDP细胞内铂类药物的浓度并改善其铂类药物不敏感性。更好的是，在给予人参皂苷Rb1加铂类药物的联合治疗后可能没有不可预测的CYP3A4介导的药代动力学相互作用。人参皂苷Rb1是A549/DDP细胞铂类药物不敏感的可能逆转剂，具有通过靶向ABCB1和Hedgehog（Hh）通路抑制两种药物泵（P-gp和PTCH1）的外输的双重功能。总的来说，本研究为开发铂类药物不敏感NSCLC的新型逆转剂奠定了基础。版权所有©2023年。Elsevier GmbH出版。

The multi-drug resistance is an inherent weakness in the chemotherapeutics of non-small cell lung cancer occurring frequently all over the world. Clinically, ginseng and Chinese medicinal prescriptions including ginseng usually used as anti-tumor adjuncts due to its characteristic of qi-invigorating, which could improve the curative effect of chemotherapy drugs and reduce their toxic side effects. Triterpenoid saponins are the crucial active ingredients in Panax ginseng, and Ginsenoside Rb1 is of the highest quantities. However, the research on the tumor drug-resistance reversal effect and mechanism of ginsenoside Rb1 is still not clear.This study aimed to systematically estimate the reversal activity of Ginsenoside Rb1 on cisplatin-insensitivity of A549/DDP cells and to reveal its prospective molecular mechanism.MTT assay were conducted to evaluate the reversal activity on cisplatin-insensitivity of A549/DDP cells of Ginsenoside Rb1in vitro, and the behavior was also studied by establishing a subcutaneous transplanted tumor model of A549/DDP in BALB/c-nu mice. In addition, P-gp ATPase activity assay, cisplatin accumulation assay, Annexin V-FITC apoptosis assay, real-time qPCR analysis and western blotting analysis were used to clarify the potential mechanism.Ginsenoside Rb1 could effectively reverse the cisplatin-resistance of A549/DDP in vitro and vivo. And after the co-treatment of Ginsenoside Rb1 plus cisplatin, the accumulation of cisplatin increased in A549/DDP cells, which was accompanied with the down-regulation of the mRNA and protein expression levels of ABCB1, SHH, PTCH1 and GLI2. Besides, the apoptosis-inducing ability of cisplatin improved by the relative regulation on the protein expression level of Bax and Bcl-2. Far more importantly, the changes of CYP3A4 mRNA and protein levels were not significant.Ginsenoside Rb1 could increase the concentration of intracellular cisplatin and improve the insensitivity for cisplatin on A549/DDP cells. Even better, there was perhaps no unpredictable CYP3A4-mediated pharmacokinetic interactions after the combination of Ginsenoside Rb1 plus cisplatin. Ginsenoside Rb1 was a probable reversal agent for the cisplatin-insensitivity of A549/DDP cells, with a bifunction of inhibiting the efflux of two drug pumps (P-gp and PTCH1) by targeting ABCB1 and Hedgehog (Hh) pathway. In general, this research laid the groundwork for the development of a new reversal agent for the cisplatin-insensitivity of NSCLC.Copyright © 2023. Published by Elsevier GmbH.